沉默ANGPTL2对氧—葡萄糖剥夺/复氧诱导的HT22细胞氧化应激及凋亡的影响

目的 探究沉默血管生成素样蛋白2(angiopoietin-like protein 2,ANGPTL2)对氧-葡萄糖剥夺/复氧(oxygen-glucose deprivation/reoxygenation, OGD/R)诱导的神经元细胞(hippocampal neuronal cell line, HT22)细胞氧化应激及凋亡的影响。方法 通过OGD/R处理建立小鼠海马HT22细胞模型。通过实时荧光定量聚合酶链反应、细胞计数试剂盒、末端标记法及试剂盒检测ANGPTL2 mRNA的含量、细胞活力、细胞凋亡、细胞乳酸盐脱氢酶(lactate dehydrogenase, LDH)水平及细胞氧化应激。通过蛋白质印迹检测各种蛋白质含量。结果 与对照组比较，模型组ANGPTL2 mRNA表达水平、LDH、凋亡率、ROS、MDA、核苷酸寡聚化结构域样受体家族3(nucleotide oligomeric domain-like receptor family 3,NLRP3)及凋亡相关斑点样蛋白(apoptosis-associated speck-like protein contain...

Effects of ANGPTL2 silencing on oxidative stress and apoptosis of HT22 cells induced by oxygen-glucose deprivation/reoxygenation

Objective To explore the effects of angiopoietin-like protein 2 (ANGPTL2) silencing on oxidative stress and apoptosis of hippocampal neuronal cell line HT22 cells induced by oxygen-glucose deprivation/reoxygenation (OGD/R).Methods The mouse HT22 cell model was established by OGD/R treatment. ANGPTL2 mRNA level, cell viability, cell apoptosis, lactate dehydrogenase (LDH) level, and cellular oxidative stress were determined by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), cell count kit, terminal labeling and kit. The levels of various proteins were detected by Western blotting.Results Compared with the control group, ANGPTL2 mRNA expression level, LDH, apoptosis rate, reactive oxygen species (ROS), malondialdehyde (MDA), nucleotide oligomeric domain-like receptor family 3 (NLRP3), apoptosis-associated speck-like protein containing a card (ASC), nuclear factor E2 related factor 2 (Nrf2) and protein expressions of heme oxygenase-1 (HO-1) were significantly increased (P＜0.05), while cell viability and super oxide dismutase (SOD) were significantly decreased (P＜0.05). Compared with OGD/R+si-con group, ANGPTL2 mRNA expression level, LDH, apoptosis rate, ROS, MDA, NLRP3 and ASC protein expression were significantly decreased in OGD/R+ Si-Con group (P＜0.05), while cell viability, and SOD as well as Nrf2 and HO-1 protein expression were significantly increased (P＜0.05).ConclusionSilencing ANGPTL2 alleviates OGD/R-induced oxidative stress and NLRP3 inflammasome mediated neuronal apoptosis by regulating Nrf2/HO-1 signaling pathway.